Three new species of Begonia endemic to the Puerto Princesa

Rubite et al. Bot Stud (2015) 56:19
DOI 10.1186/s40529-015-0099-1
Open Access
RESEARCH
Three new species of Begonia endemic
to the Puerto Princesa Subterranean River
National Park, Palawan
Rosario Rivera Rubite1, Mark Hughes2, Patrick Blanc3, Kuo‑Fang Chung4, Hsun‑An Yang5, Yoshiko Kono5,
Grecebio J D Alejandro6, Llogene B De Layola1, Arthur Gregory N Virata1 and Ching‑I Peng5*
Abstract Background: Begonia is a mega-diverse genus of flowering plants prone to generating micro-endemic species,
especially on limestone habitats. During fieldwork in the Puerto Princesa Subterranean River National Park, Palawan
(Philippines), three species were encountered which did not match any previously described from the region.
Results: Following morphological, anatomical, molecular phylogenetic and cytological investigation a hypothesis of
three new species is supported. The three new species belong to a clade endemic to Palawan and Borneo.
Conclusions: The limestone habitats in the Puerto Princesa Subterranean River National Park environs support a
unique flora. The description of three new species from a small area within the park demonstrates how much remains
to be discovered there, and the importance of its continued protection.
Keywords: Limestone, Endemic, New species, Conservation
Background
The Philippines is a biodiversity hotspot, with an exceptional concentration of endemic species undergoing
exceptional loss of habitat (Myers et al. 2000). Some of
the most extensive areas of intact forest in the country
are found in Palawan, where there are reasons for optimism due to the implementation of conservation through
devolved local government and civil society groups (Posa
et al. 2008). Puerto Princesa Subterranean River National
Park (PPSRNP) is managed by the City Government of
Puerto Princesa in partnership with the Department of
Environment and Natural Resources (DENR), and is the
first national park in the Philippines to be managed at
this level. It was designated a World Heritage Site in 1999
(World Heritage Commitee 2000), and voted as one of
the New 7 Wonders of Nature, boosting tourism as much
as 300% (Fitzgerald 2011, 2012). Palawan as a whole is
designated by UNESCO as a Biosphere Reserve, and by
*Correspondence: [email protected]
5
Biodiversity Research Center, Academia Sinica, Taipei 115, Taiwan
Full list of author information is available at the end of the article
Birdlife International as an Important Bird and Biodiversity Area (BirdLife International 2015).
The PPSRNP contains a spectacular karst limestone
landscape and eight different forest formations; forest
on ultramafic soil, forest on limestone soil, montane forest, freshwater swamp forest, lowland evergreen tropical
rainforest, riverine forest, beach forest, and mangrove
forest. This mix of forest types and substrates means the
PPSRNP harbours a very rich flora, but much remains to
be discovered. During recent fieldwork in the PPSRNP
we found three new species of Begonia within a distance
of only a few hundred meters, adding these distinct species to the 14 known Begonia species already recorded
from Palawan (Hughes and Coyle 2009; Hughes et al.
2010, 2011). Based on morphology, chromosome cytology and molecular phylogenetic analysis we place the
three new species in Begonia sect. Baryandra. Although
the species occur very close to one another at altitudes of
less than 50 m, they grow in mutually exclusive habitats.
The first species Begonia taraw C.-I Peng, R. Rubite & M.
Hughes is located around the mouth of the underground
river, growing tenaciously on the vertical limestone
© 2015 Rubite et al. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License
(http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium,
provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license,
and indicate if changes were made.
Rubite et al. Bot Stud (2015) 56:19
cliffs. Nearby in karst forest growing in rock crevices is
the second species Begonia hughesii R. Rubite and C.-I
Peng. The third species, Begonia tagbanua M. Hughes,
C.-I Peng & R. Rubite, grows on clay soil banks inside the
forest.
Methods
Chromosome cytology
Somatic chromosomes of B. taraw (Ching-I Peng 23463,
23464), B. hughesii (Ching-I Peng 23466, 23475), B. tagbanua (Ching-I Peng 23471) were examined using root
tips. The procedures of pre-treatment, fixation and staining for chromosome observations followed Peng et al.
(2014). Voucher specimens are deposited in HAST.
Cryo scanning electron microscopy
Fresh leaves of B. taraw, B. hughesii and B. tagbanua were
dissected and attached to a stub. The samples were frozen
with liquid nitrogen slush, then transferred to a sample
preparation chamber at −160°C and etched for 15 min at
−85°C. After etching, the temperature reached −130°C
for sample fracturing and coating. After coating, the samples were transferred to the SEM chamber and observed
at −190°C with a cryo scanning electron microscope (FEI
Quanta 200 SEM/Quorum Cryo System PP2000TR FEI).
Phylogenetic analyses
A total of 33 species from Begonia sect. Baryandra
was sampled, including the 3 new species from Palawan (each represented by two individuals) and a further 5 from Palawan (Begonia blancii, B. cleopatrae,
Begonia suborbiculata, B. wadei, B. woodii). In addition
three species of Begonia sect. Reichenheimia were sampled, with two species of Begonia sect. Coelocentrum
as an outgroup based on the tree topology in Thomas
et al. (2012). Vouchers and genbank accession numbers
are in Additional file 1: Table S1. Begonia sect. Baryandra has recently been recircumscribed and confirmed
as monophyletic (Rubite et al. 2013). Total genomic
DNA was extracted from young leaves and buds using
DNeasy Plant Mini Kits (Qiagen, USA). Four chloroplast
non-coding regions (ndhA intron, ndhF–rpl32 spacer,
rpl32–trnL spacer, trnC–trnD spacer) were amplified as
in Hughes et al. (2015) with the PCR primer sequences
from Demesure et al. (1995) and Thomas et al. (2011).
Forward and reverse reads for all regions were assembled
and aligned using Geneious Pro (Biomatters, New Zealand), with the alignments being subsequently manually
edited in BioEdit 7.1.3 (Hall 1999); inversions were offset and the final alignment was 6905 bases long, with 39
bases excluded due to alignment uncertainty. The optimal model of DNA sequence evolution was assessed with
jModeltest 2.1.3 (Posada et al. 2012) using the Aikake
Page 2 of 14
Information Criterion (AIC) to estimate the model with
the closest fit to the data. The GRT + G + I model was
the most probable (AIC weight of 0.98). Bayesian phylogenetic analysis was carried out using MrBayes 3.2.1
(Ronquist et al. 2012), treating the dataset as a single partition. The analysis consisted of two runs with four chains
each, run for 10,000,000 generations with a sample tree
taken every 1,000. The effective sample size was >5,600
for all model parameters, and the average standard deviation of split frequencies was 0.0016. The proportion
of successful state exchanges between adjacent chains
ranged between 0.62 and 0.68 for both runs. Convergence between runs and attainment of stationarity was
runs was further comfirmed by examining parameter distrubutions in Tracer v1.6 (Rambaut et al. 2014). The first
25% of sampled trees were discarded as burn-in, and the
remainder summarised as a maximum clade credibility
tree visualised using FigTree v.1.4.0 (Rambaut 2009).
Results
Chromosome cytology
Recently, Philippine Begonia species belonging to section Diploclinium were transferred to section Baryandra, based on molecular analysis (Rubite et al. 2013).
Among 47 species in sect. Baryandra, chromosome
numbers were previously reported for five species,
namely B. blancii (2n = 30, Hughes et al. 2011); B. fenicis
(2n = 26, Oginuma and Peng 2002; 2n = 56, Kokubugata
and Madulid 2000); B. parva (2n = 36 + 2f, Legro and
Doorenbos 1969); B. nigritarum (2n = 44, Doorenbos
et al. 1998); and B. suborbiculata (2n = 30, Hughes et al.
2011). Somatic chromosomes at metaphase of the three
new species are reported here for the first time: 2n = 28
for Begonia taraw (Figure 1a); 2n = 30 for B. hughesii
(Figure 1b) and B. tagbanua (Figure 1c).
Leaf anatomy and vestiture
Begonia taraw
Adaxial surface nearly glabrous, epidermis cells slightly
bullate (Figure 2a, b); cross section ca. 820 μm thick;
epidermis single-layered on both surfaces, hypodermis
2-layered; palisade parenchyma cells 1-layered; spongy
parenchyma cells ca. 3-layered (Figure 2b); abaxial surface with glandular hairs and multiseriate trichomes, stomata in clusters of 3–7, helicocytic, flat, subsidiary cells 3
(Figure 2c).
Begonia hughesii
Adaxial surface with glandular trichomes (Figure 2d);
cross section ca. 870 μm thick; epidermis single-layered on both surfaces, hypodermis 2-layered; palisade
parenchyma cells 1-layered; spongy parenchyma cells
ca. 4-layered (Figure 2e); abaxial surface with glandular
Rubite et al. Bot Stud (2015) 56:19
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Figure 1 Somatic chromosomes at metaphase of Begonia. a B. taraw (2n = 28 [0.9–1.6 µm], Peng 23464); b B.hughesii (2n = 30 [1.0–2.1 µm], Peng
23466); c B. tagbanua (2n = 30 [0.9–2.0 µm], Peng 23471).
Figure 2 Leaf SEM microphotographs of Begonia. a–c Begonia taraw; d–f B. hughesii; g–i B. tagbanua; a, d, g, adaxial surface; b, e, h, cross section; c,
f, i, abaxial surface.
Rubite et al. Bot Stud (2015) 56:19
trichomes, stomata single or 2–5 clustered, helicocytic,
flat, subsidiary cells 3 (Figure 2f ).
Begonia tagbanua
Adaxial surface with conoidal cells and glandular trichomes (Figure 2g, h); Cross section ca. 490 μm thick;
epidermis single-layered on both surfaces, hypodermis
1-layered; palisade parenchyma cells 1-layered; spongy
parenchyma cells ca. 2-layered (Figure 2h); abaxial surface with glandular trichomes, stomata 2–4 clustered,
helicocytic, flat, subsidiary cells 3 (Figure 2i).
Phylogenetic relationships
The three new species B. hughesii, B. tagbanua and
B. taraw are strongly supported as being members of
Begonia sect. Baryandra, and belong to a clade mostly
endemic to Palawan (Figure 3). Begonia hughesii is not
monophyletic according to its chloroplast genotype,
whereas B. tagbanua and B. taraw are supported as
monophyletic whilst having some within-species polymorphism as shown by the non-zero branch lengths. The
two samples of Begonia hughesii were taken from different localities; sample 1 (Peng 23466) was collected in littoral forest near Sabang, and sample 2 (Peng 23475) was
collected at Ugong Rock.
Taxonomic treatment
1. Begonia taraw C.-I Peng, R. Rubite & M. Hughes, sp.
nov. § Baryandra (Figures 4, 5)
TYPE: PHILIPPINES. Palawan, Puerto Princesa, Puerto
Princesa Subterranean River National Park, elev. ca. 5 m,
10°12′1″N, 118°55′32″E, 2 Nov. 2011, Ching-I Peng 23464,
with Kuo-Fang Chung, Chien-I Huang, Rosario Rubite
(holotype PNH, isotype E, HAST).
Begonia taraw can be differentiated from the only other
peltate species in Palawan, Begonia gutierrezii, in having
longer petioles which are sparsely lanate with appressed
hairs (not erect 4 mm long hairs), short internodes on the
rhizome (not stoloniferous and elongate), larger leaves,
glabrous stipules, and a conspicuous ring of fused fleshy
hairs at the base of most petioles.
Lithophytic rhizomatous herb 20–35 cm tall. Stem
11–16 mm diameter, glabrous, internodes 5–10 mm
long.
Stipules
broadly
triangular,
asymmetric,
13–15 × 15–17 mm, keeled, acuminate, entire, glabrous,
persistent, maroon becoming dark brown and papery with
age. Leaves on erect petioles; petioles 13–20 cm long, red at
the base, greener apically, terete, 5–7 mm in diameter at the
base becoming more slender apically, sparsely lanate when
young, the hairs appressed, subglabrous with age, the hairs
rubbing off easily, broad-based 10 mm long fleshy red hairs
fused into a ring at the base of the petiole; lamina peltate,
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petiole inserted 3–3.5 cm from midrib, distinctly coriaceous, not variegated, in various colour forms on different
plants from pale to very dark matt green above, whitish to
pinkish green below, broadly ovate, 6–8 × 9–10 cm, upper
surface mostly glabrous, sometimes with a few scattered
2 mm fine long brown hairs between the veins, the pale
eye above the petiole attachment with a few hairs; lower
surface sparsely lanate on the veins and lamina; venation
palmate, main veins 9, pink or green, slightly prominent
beneath, flat above; apex obtuse; margin very shallowly
sinuate-dentate; edges reddish becoming green when
mature; stomata in clusters of 3–7. Inflorescence erect,
40–58 cm long, bisexual, male and female flowers open
at the same time as the inflorescence matures, cymose,
each unit consisting of a central male flower with a lateral
female and lateral further unit, branching up to 6 times,
with 40–80 flowers in total, not mass-flowering; primary
peduncle 30–45 cm long, 5–7 mm diameter at the base
becoming more slender apically, base is reddish becoming
green at the upper third part, sparsely lanate when young,
subglabrous with age, the hairs rubbing off easily, secondary peduncles 1.5–4 cm long, subglabrous; bracts broadly
rounded-triangular, conduplicate, ca. 10 × 12 mm, entire,
glabrous, deciduous, overlapping, translucent pale green
or pinkish green. Male flowers pedicel 8–15 mm long, glabrous; tepals 4, uniformly white or pink, entire, glabrous;
outer 2 oblong, 13–17 × 10–13 mm; inner 2 obovate,
folded and retuse, 11–14 × 8–9 mm; androecium with ca.
50 stamens, sessile; filaments united at the base, unequal,
1.25–2 mm long, longer filaments found in the centre of
the androecium; anthers yellow, 1.75 mm long, oblongoblanceolate, slits lateral tending to unifacial towards the
apex, running for 3/4 the length of the anther, connective
extended. Female flowers pedicel 20–30 mm long, glabrous,
with 2 min fleshy 0.5 mm long hair-like bracteoles spaced
1–2 mm below the ovary; ovary 12 × 20 mm including
wings, green or pink; capsule ca. 9 × 6 mm, 3-locular, placentae bifid; wings 3, unequal, larger wing ca. 9 × 14 mm,
shallowly angular to rounded, flat (not cucullate), smaller 2
wings ca. 11 × 4 mm, truncate; tepals the same as the male
flowers; styles 3, bifid, stigmatic surface spirally twisted.
Fruit drying pale brown, the same size and shape as the
ovary. Somatic chromosome number, 2n = 28.
Distribution, habitat and ecology Begonia taraw is currently known only from the type locality and a further
field observation from Lions Cave ca. 500 m away. The
species grows on vertical limestone cliffs semi-shaded
by broadleaf forest at the entrance of the Puerto Princesa
underground river. The species was flowering and fruiting when collected in early November. In cultivation in
the greenhouse of Academia Sinica in Taipei, Taiwan, it
flowered and fruited from July to December.
Rubite et al. Bot Stud (2015) 56:19
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Figure 3 Phylogeny of Begonia sect. Baryandra. Generated from a Bayesian analysis of chloroplast DNA sequence data, the 50% majority-rule con‑
sensus tree is shown. The figures at the nodes are clade support values (posterior probabilities).
Etymology The species is named after the local word “taraw”
which means karst limestone in Tagbanua language.
Additional specimen examined (paratype) PHILIPPINES.
Palawan, Puerto Princesa Subterranean River National
Park, elev. ca. 5 m, 10°12′1″N, 118°55′32″E, 2 Nov. 2011,
Ching-I Peng 23463, with Kuo-Fang Chung, Chien-I
Huang, Rosario Rubite (HAST).
Rubite et al. Bot Stud (2015) 56:19
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Figure 4 Begonia taraw C.-I Peng, R. Rubite, & M. Hughes. a Habit; b showing the base of petiole; b’, stipule; b”, fleshy hairs fused into a ring; c
bract; d, d’ staminate flower; e, e’ stamen; f, f’ carpellate flower; g, g’, g” style and stigma; h capsule; i cross section of ovary. All from C.-I Peng 23463
(HAST). Line drawing by Che-Wei Lin.
Proposed IUCN red list category VUD2.The species is a
rare, very narrow endemic, and as such has an area of occupancy and extent of occurrence under the threshold for
Critically Endangered, (IUCN 2012). However, none of the
other criteria, based on number of locations and decline or
fluctuation in range or population apply. B. taraw is best
considered as belonging to the Vulnerable category, under
VUD2, a category often applied to naturally rare and narrowly endemic taxa which, although not under immediate
threat, have such a narrow range that unpredictable localised habitat degradation puts them at risk of rapidly fulfilling the criteria for Critically Endangered or Extinct.
2. Begonia hughesii R. Rubite & C.-I Peng, sp. nov. § Baryandra (Figures 6, 7)
TYPE: PHILIPPINES. Palawan Island, Palawan Province,
Puerto Princesa City, Puerto Princesa Subterranean River
Rubite et al. Bot Stud (2015) 56:19
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Figure 5 Begonia taraw C.-I Peng, R. Rubite & M. Hughes. a Habit and habitat; b inflorescence; c petiole; d leaf adaxial surface; e leaf abaxial surface,
red type; f young leaf, stipules and fleshy hairs fused into a ring at the base of the petiole; g bracts and young inflorescence; h staminate flower, face
view; i 4-tepaled carpellate flower, face view; j 5-tepaled carpellate flower, face view; k staminate flower, side view; l carpellate flower, side view; m
cross section of ovary. All from C.-I Peng 23463 (HAST) except E & J from C.-I Peng 23464 (HAST).
National Park, elev. ca. 5 m, 10°12′1″N, 118°55′32″E, 2
Nov. 2011, Ching-I Peng 23466, with Kuo-Fang Chung,
Chien-I Huang, Rosario Rubite (holotype PNH, isotype
HAST).
Begonia hughesii is most similar to B. acclivis, but differs in having leaves which are glabrous above (not with
scattered 2 mm long hairs) and uniform bright green (not
markedly variegated), and dimorphic stipules which are
either hairy on the keel only or hairy all over the abaxial
surface.
Lithophytic rhizomatous herb ca. 20 cm tall. Stem
5–10 mm thick, tomentose (2–3 mm brown hairs) becoming glabrous in parts with age, internodes 3–8 mm long.
Stipules triangular, asymmetric, 10–12 × 9–11 mm,
Rubite et al. Bot Stud (2015) 56:19
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Figure 6 Begonia hughesii R. Rubite & C.-I Peng. a Habit; b stipule; c bract; d, d’ staminate flower; e, e’ stamen; f, f’ carpellate flower; g, g’, g” style
and stigma; h capsule; i cross section of ovary. All from C.-I Peng 23466 (HAST). Line drawing by Che-Wei Lin.
heteromorphic, alternately either with brown hairs on
the keel only, or tomentose all over abaxially, hairs brown,
2–3 mm long, keeled, keel extending at the tip for ca. 2 mm,
ring of hairs at base of petiole absent. Leaves on erect petioles; petioles 6–15 cm long, 4–6 mm in diameter, terete,
red, tomentose even when mature and with 2 hair types,
one linear, the other thicker towards the base; lamina basifixed, thick and fleshy in life, drying opaque and stiff, ovate,
7–10 × 5–7.5 cm, apple-green above, pale green below,
glabrous above except for minute pubescence near the
petiole attachment, below hairy on the veins only with ca.
2 mm long hairs, venation palmate, main veins 7; margin
maroon, broadly and shallowly undulate, dentate-sinuate,
ciliate with pink hairs (1–1.5 mm); tip shortly acuminate;
stomata in clusters of 2–5. Inflorescence erect, ca. 20 cm
long, bisexual, male and female flowers open at the same
time as the inflorescence matures, cymose, branching 4
times; primary peduncle ca. 15 cm long, with scattered
hairs (much less dense than on the petioles), bracts small,
ovate, 1.5–2 × 1.5–2 mm, with minute scattered glandular
Rubite et al. Bot Stud (2015) 56:19
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Figure 7 Begonia hughesii R. Rubite & C.-I Peng. a Habit and habitat; b cultivated plant at anthesis; c inflorescence; d rhizome, stipules and petioles;
e bracts and young inflorescence; f carpellate flower, face view; g carpellate flower, side view; h leaf adaxial surface; i leaf abaxial surface; j capsule;
k cross section of ovary. All from C.-I Peng 23466 (HAST).
Rubite et al. Bot Stud (2015) 56:19
hairs. Male flowers: pedicel 7–10 mm long, with sparse
minute glandular hairs; tepals 4, uniformly very pale pink
or white, entire, with a few minute glandular hairs adaxially; outer 2 sub-orbicular 8–9 × 6–7 mm; inner 2 obovate, slightly retuse, 6 × 3 mm; androecium with ca. 40
stamens, sessile; filaments free, unequal, 1–1.75 mm
long, longer filaments found in the centre of the androecium; anthers yellow, 1.25 mm long, oblong-oblanceolate,
slits lateral becoming unifacial at the apex; connective
extended. Female flowers: pedicel 7–10 mm long, with
sparse glandular hairs; ovary 10 × 14 mm including wings,
pink; capsule 9 × 5 mm, two-locular, placentae bifid; wings
3, unequal, larger wing cucullate, smaller wings rounded;
tepals the same as in the male flowers; styles 3, stigmatic
surface spirally twisted. Fruit pedicel 7–12 mm, drying
pale brown, 9–14 × 14–18 mm, recurved with the 2 lateral wings (9–14 × 3–4 mm, round) forming a splash cup,
the larger wing markedly cucullate, 9–14 × 7–9 mm, apex
truncate. Somatic chromosome number, 2n = 30.
Distribution, habitat and ecology Begonia hughesii herbarium material has been collected from the type locality
near the entrance to the Underground River and on Ugong
Rock near Sabang and on limestone outcrops at the base of
Mt. St. Paul. Further photographic evidence reveal records
from the other side of the Babuyan River at PPURNP and
within the forest on Cleopatra’s Needle (Jonah Van Beijnen, pers. comm.). The species grows on eroded limestone
in semi-shade under broadleaf forest, and was flowering
and fruiting when collected in early November. In cultivation in the greenhouse of Academia Sinica in Taipei, Taiwan, it flowered and fruited from July to December.
Etymology The species epithet is named after Dr Mark
Hughes of the Royal Botanical Garden Edinburgh, who
studies extensively Southeast Asian begonias including
Philippine Begonia.
Additional specimens examined (paratypes) PHILIPPINES. Palawan, Sabang, Ugong Rock, 10°5′13″N,
118°51′16″E, elev. ca. 60 m, in limestone outcrops, semishaded, locally abundant, 2 Nov. 2011, Ching-I Peng
23475, with Kuo-Fang Chung, Chien-I Huang, Rosario
Rubite (HAST); Palawan, Mt. St. Paul, elev. 300–600 m,
slope forest on karst limestone, 03-05 May 2012, Hughes
& Tandang MH1726 (HAST); Palawan, Puerto Princesa
Subterranean National Park, Sabang, jungle trail between
CPS and underground river, elev. 50 m, 29 May 2012, M.
Hughes, C. Puglisi, D. Tandang & Julius CP305 (E, PNH).
Proposed IUCN Red list category LC. The species has a
fairly wide distribution within the PPURNP and surrounding forests on Mt St Paul and Cleopatra’s Needle, and providing these areas remain well managed B.
hughesii can be considered as Least Concern.
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3. Begonia tagbanua M. Hughes, C.-I Peng & R. Rubite,
sp. nov. § Baryandra (Figures 8, 9)
TYPE: PHILIPPINES. Palawan, Puerto Princesa, Puerto
Princesa Subterranean River National Park, elev. ca.
35 m, 10°12′1″N, 118°55′32″E, 2 Nov. 2011, Ching-I Peng
23471, with Kuo-Fang Chung, Chien-I Huang, Rosario
Rubite (holotype PNH, isotype A, E, HAST, MO).
Begonia tagbanua is most similar to B. suborbiculata,
but differs in having less succulent leaves which are
rugolose (not smooth), sparsely puberulous above (not
glabrous), and fruits which are 3-winged (not 5-winged).
Terrestrial or lithophytic rhizomatous herb ca. 10 cm
tall. Stem 6–8 mm in diameter, with sparse 1–2 mm
brown hairs becoming glabrous with age, internodes
4–5 mm long. Stipules triangular, 14–16 × 4–5 mm,
greenish to pink, with sparse hairs, keel present, apex
fimbriate, margin entire. Leaves on petioles relaxed
against the substrate; petiole 11–13 cm long and 4–6 mm
in diameter, terete, succulent, maroon densely covered with brown straight hairs (1–2 mm), simple 2 mm
long red hairs in a ring at the base of the petiole; blade
ovate to suborbicular; 2 colour forms: one is adaxial surface dark green with brown patches at interveins and
abaxial surface maroon, while another is adaxial surface plain apple green and abaxial surface light green;
adaxial surface has white straight hairs (2–3 mm) arising from a green or brown dot while abaxial surface has
white straight hairs (2–3 mm) at veins and shorter hairs
(1–1.5 mm) at interveins, base cordate, lobes rounded,
sinus overlap 1 cm, apex obtuse, margin shallowly undulate, denticulate, ciliate with brown straight hairs (1 mm),
primary veins 7–8, stomata in clusters of 2–4. Inflorescence erect, 22–31 cm long, axillary, arising directly from
the rhizome, bisexual, male and female flowers open
at the same time as the inflorescence matures, 4 times
dichotomously branching; peduncle pink 16–23 cm long,
3–4 mm in diameter, erect, with sparse light brown hairs
(1–1.5 mm); bracts rounded-triangular, 4 × 2 mm, glabrous; pedicels pink 1.6–1.8 cm erect to ascending in
staminate flowers, 1.5 cm ascending to horizontal in carpellate flowers. Male flowers: pedicel ca. 10 mm, glabrous;
tepals 4, pink, glabrous, outer pair orbicular with 10–12
veins; 11–12 × 10–11 mm, inner pair obovate folded
and retuse, 8–13 × 7–8 mm; stamens 30–40; filaments
1 mm long, united at the base; anthers yellow, rounded,
0.5 mm long. Female flowers: pedicel ca. 15 mm, ovary
10 × 14 mm including wings, pink; capsule 9 × 4 mm,
two-locular, placentae bifid; wings 3, unequal, larger wing
cucullate, smaller wings rounded; tepals 4, pink, glabrous, outer pair suborbicular 13–16 × 10–12 mm; inner
pair obovate, folded, retuse 11–13 × 4–6 mm; styles 3,
stigmatic surface spirally twisted. Fruit pedicel 1–1.3 cm;
Rubite et al. Bot Stud (2015) 56:19
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Figure 8 Begonia tagbanua M. Hughes, C.-I Peng & R. Rubite. a Habit; b indumentum on leaf adaxial surface; c stipule; d Bract; e, e’, staminate
flower; f, f’ stamen; G,G’, carpellate flower; g, h’, h”, style and stigma; i Capsule; j cross section of ovary. All from C.-I Peng 23471 (HAST). Line drawing
by Che-Wei Lin.
capsule rounded in outline, glabrous, 8–12 × 18–20 mm
unequally 3-winged, adaxial wing strongly cucullate
shorter than the capsule 7–10 × 8–11 mm; lateral wings
8–12 × 5–6 mm forming a splash cup. Somatic chromosome number, 2n = 30.
found growing on steep banks of bare soil such as can be
found at the base of tree roots. The species was flowering
and fruiting when collected in early November. In cultivation in the greenhouse of Academia Sinica in Taipei, Taiwan, it flowered and fruited from July to December.
Distribution, habitat and ecology Begonia tagbanua is
currently known from the type locality in PPSRNP, and
from the foothills at the base of Mt. St. Paul at ca. 200 m
altitude (M. Hughes field observation). In PPSRNP it
grows quite abundantly along trail sides in bare soil, under
semi-shaded broadleaf forest. Away from trails it can be
Etymology The species is named after the Tagbanuas, one
of the indigenous peoples of the type locality.
Additional specimens examined (paratypes) PHILIPPINES. Palawan, Puerto Princesa, Puerto Princesa Subterranean River National Park, elev. ca. 35 m,
Rubite et al. Bot Stud (2015) 56:19
Page 12 of 14
Figure 9 Begonia tagbanua M. Hughes, C.-I Peng & R. Rubite. a Habit and habitat; b cultivated plant at anthesis; c carpellate flower, side view; d
carpellate flower, face view; e staminate flower, face view; f staminate flower, side view; g stipules; h leaf adaxial surface, showing indumentum; i
capsule; j cross section of ovary. All from C.-I Peng 23471 (HAST).
10°12′1″N, 118°55′32″E, 2 Nov. 2011, Ching-I Peng
P23472, with Kuo-Fang Chung, Chien-I Huang, Rosario
Rubite (HAST); Palawan, Puerto Princesa Subterranean
National Park, Sabang, jungle trail between CPS and
underground river, elev. 50 m, 10°11′53″N, 118°55′5″E,
29 May 2012, M. Hughes, C. Puglisi, D. Tandang & Julius
CP301 (E, PNH).
Proposed IUCN Red list category LC. In common with B.
hughesii and B. taraw, B. tagbanua is a narrow endemic
Rubite et al. Bot Stud (2015) 56:19
restricted to the PPSRNP environs. However it is not an
obligate lithophyte, and can colonise recently dug clay
soil banks by trail sides, and indeed the largest population
observed was along the coastal trail toward the subterranean river cave entrance. Hence the appropriate conservation category for the species is Least Concern.
Discussion
The variety of habitats in terms of forest types and substrates, and in particular the variety of limestone habitats,
may play a part in promoting species richness at this relatively low altitude for Begonia (Chung et al. 2014). Further
exploration towards the karst summit of the nearby Mount
St. Paul did not yield any more Begonia species, contrary
to what we would expect for this predominantly montane
genus. The phylogenetic results show that all three species
harbour chloroplast polymorphism, which is congruent
with Begonia species existing as populations with a high
degree of genetic isolation (Hughes and Hollingsworth
2008; Twyford et al. 2014). Both B. tagbanua and B. taraw
resolved as monophyletic in the chloroplast phylogeny,
whereas B. hughesii appears as polyphyletic. Natural hybrids
have been reported a number of times in Begonia (Peng and
Chiang 2000; Peng and Ku 2009; Peng et al. 2010) and it is
possible that B. hughesii and B. tagbanua share a common
chloroplast lineage due to a past hybridisation event.
Conclusions
The study further highlights the importance of the Puerto
Princesa Subterranean River National Park as a remarkable World Heritage Site, with its combination of unique
geology and endemic flora. The presence of three narrowly
endemic and rare species in the most heavily visited part of
the reserve means continued care must be taken to ensure
trails are well maintained and that they are respected by
visitors, and that visitor numbers are managed appropriately. Banks by trail sides can provide an ideal habitat for
Begonia, particularly if the understory vegetation along the
trail is left as intact as possible to provide shade and preserve the micro-climate. Ex-situ conservation collections
also have a role to play in ensuring a secure future for the
three species described here, giving the potential for reintroduction if a chance event such as drought or typhoon
should destroy or drastically reduce the wild populations.
The fact that three new species were found in such close
proximity demonstrates how much remains to be discovered in the PPSRNP, and how even small scale forest
disturbance could result in extinctions. The study supports the formation of a larger forest reserve encompassing both PPSRNP and the adjacent Cleopatra’s Needle
Mountain Range, which also has two recently discovered
endemic Begonia (Hughes et al. 2010), and doubtless
many other endemic plant species awaiting description.
Page 13 of 14
Additional file
Additional file 1. Voucher specimen details and genbank accession
numbers for the DNA sequences used in the phylogenetic analysis.
Authors’ contributions
RR, MH, PB, GA, KFC and CIP carried out field observations of the new taxa and
conceived of the study. RR, HAY, LL and AV drafted the species descriptions.
YK carried out the Chromosome cytology. HAY carried out the cryo scanning
electron microscopy studies. MH carried out the molecular phylogenetic
analysis. All authors read and approved the final manuscript.
Author details
1
Department of Biology, College of Arts and Sciences, University of the
Philippines Manila, Padre Faura, Manila 1000, Philippines. 2 Royal Botanic
Garden Edinburgh, 20a Inverleith Row, Edinburgh EH3 5LR, UK. 3 CNRS, 3 rue
Michel‑Ange, 75794 Paris, France. 4 School of Forestry and Resource Conserva‑
tion, National Taiwan University, Taipei 106, Taiwan. 5 Biodiversity Research
Center, Academia Sinica, Taipei 115, Taiwan. 6 College of Science and Research
Centre for the Natural and Applied Sciences, University of Santo Tomas,
España, Manila 1015, Philippines.
Acknowledgements
We thank the Palawan Council for Sustainable Development, the Palawan
Protected Areas Management Board, Palawan City Council and the Depart‑
ment of Environment and Natural Resources for granting access and research
permits; Chien-I Huang (HAST) for assistance in the field trip; Guan-Bin Chen
for cryo-SEM studies, Che-Wei Lin (TAIF) for the line drawings, and Jonah
Van Beijnen for comments on the manuscript. The curators of BM, E, HAST,
K, PNH herbaria are thanked for providing access to specimens. The Royal
Botanic Garden Edinburgh is supported by the Scottish Governments Rural
and Environment Science and Analytical Services Division. This research was
supported in part by grants from the Royal Society of Edinburgh Bilateral
Programme, University of the Philippines System Enhanced Creative Work and
Research Grant (ECWRG 2014-09), Ministry of Science and Technology, Taiwan
(NSC 101-2621-B-001-003-MY3; NSC 98-2621-B-001-002-MY3) and Academia
Sinica, Taiwan.
Compliance with ethical guidelines
Competing interests
The authors declare that they have no competing interests.
Received: 29 April 2015 Accepted: 1 July 2015
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